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PMID: 165485 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Affinity chromatography of viral DNA polymerases on pyran-sepharose.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 72 ·No. 3 ·1975-03-00 ·Pages 1142-6

Chirikjian JG, Rye L, Papas TS

Abstract

Pyran covalently linked to cyanogen bromide-activated Sepharose has been shown to be an effective affinity matrix for several viral DNA polymerases. Differential salt elution of viral compared with cellular polymerases, as well as substrate elution, suggests the affinity nature for the matrix. Unlike some other affinity systems described, pyran-Sepharose is totally resistant to nuclease digestion and is stable at 4 degrees for several months. DNA polymerases isolated from several viruses by detergent treatment were recovered in good yield. Analysis of iodinated proteins by sodium dodecyl sulfate-gel electrophoresis revealed that the DNA polymerase of avian myeloblastosis virus found in crude preparations of the virus could be purified nearly to homogeneity by a single passage through the column. These results suggest that pyran-Sepharose is an effective affinity column that is potentially adaptable as part of a general purification procedure for viral DNA polymerases.

MeSH Terms
Animals Avian Myeloblastosis Virus/enzymology Avian Sarcoma Viruses/enzymology Cell Line Chick Embryo Chromatography, Affinity DNA Nucleotidyltransferases/isolation & purification Leukemia Virus, Feline/enzymology Mammary Tumor Virus, Mouse/enzymology Mice Oncogenic Viruses/enzymology RNA Viruses/enzymology Rauscher Virus/enzymology
Chemicals
DNA Nucleotidyltransferases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Chirikjian J G
Rye L
Papas T S
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25 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.72.3.1142
Published
1975-03-00
Pages
1142-6
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC432482
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.72.3.1142 PMC Full Text View journal details
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